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Organ Culture
When anesthetized, the slugs were invariably in a fully extended state,
often with the penial sac everted. The slug slowly recovers from the anesthetized state and appears normal after 20-30 min.
Dissection 1. Place the slug in a dissection tray fllied with Chiarandini's physiological
solution.
2. Dissect out very young gonads from 8-1 O-day-old slugs or the ovotestes
from stage 2 animals in Chiarandini's physiological solution, without
bicarbonate and containing penicillin G (500 IV/ml) and streptomycin (25 fig/ml), placed in a petri dish.
Culture 1. Transfer the gonads into a culture vessel.
set-up 2. Culture the gonads in gelatinized culture medium.
Results
According to Wattez (I 978), infantile or juvenile gonads can be cultured
for 3 weeks. When the gonads are cultured in isolation in a medium devoid of hormone, there is a tendency for an infantile or juvenile gonad to
progress toward the female line. The cephalic complex promotes maintenance of the male line. The optic tentacles exert an inhibitory effect on
female differentiation during the infantile and juvenile stages, but are
ineffective during the female phase. The brain favors oocyte growth, more
especially in adults of late male and female phases.
Subprotocol 2
Ovaries of Cuttlefish (Cephalopoda)
Materials
Equipment - Dissecting tray
- Scissors
- Forceps
- Culture vessels
- Pasteur pipettes
Solutions - Durchon and Schaller's medium (see Appendix 1)
Animals - Cuttlefish Sepia officinalis
Procedure
Sterilization 1. Make a longitudinal incision on the ventral side of the animal.
2. Irradiate the opened cavity with a UV ray.
Dissection 1. Excise the ovaries and optic gland (Fig. 1).
Organ Culture
When anesthetized, the slugs were invariably in a fully extended state,
often with the penial sac everted. The slug slowly recovers from the anesthetized state and appears normal after 20-30 min.
Dissection 1. Place the slug in a dissection tray fllied with Chiarandini's physiological
solution.
2. Dissect out very young gonads from 8-1 O-day-old slugs or the ovotestes
from stage 2 animals in Chiarandini's physiological solution, without
bicarbonate and containing penicillin G (500 IV/ml) and streptomycin (25 fig/ml), placed in a petri dish.
Culture 1. Transfer the gonads into a culture vessel.
set-up 2. Culture the gonads in gelatinized culture medium.
Results
According to Wattez (I 978), infantile or juvenile gonads can be cultured
for 3 weeks. When the gonads are cultured in isolation in a medium devoid of hormone, there is a tendency for an infantile or juvenile gonad to
progress toward the female line. The cephalic complex promotes maintenance of the male line. The optic tentacles exert an inhibitory effect on
female differentiation during the infantile and juvenile stages, but are
ineffective during the female phase. The brain favors oocyte growth, more
especially in adults of late male and female phases.
Subprotocol 2
Ovaries of Cuttlefish (Cephalopoda)
Materials
Equipment - Dissecting tray
- Scissors
- Forceps
- Culture vessels
- Pasteur pipettes
Solutions - Durchon and Schaller's medium (see Appendix 1)
Animals - Cuttlefish Sepia officinalis
Procedure
Sterilization 1. Make a longitudinal incision on the ventral side of the animal.
2. Irradiate the opened cavity with a UV ray.
Dissection 1. Excise the ovaries and optic gland (Fig. 1).